LINC01094 promotes human nasal epithelial cell epithelial-to-mesenchymal transition and pyroptosis via upregulating HMGB1.

BACKGROUND: Excessive epithelial-to-mesenchymal transition (EMT) of nasal epithelial cells (NECs) play a prominent role in chronic rhinosinusitis with nasal polyps (CRSwNP) pathogenesis. Long intergenic non-coding RNA 01094 (LINC01094) was previously reported to be overexpressed in CRSwNP, while the regulatory mechanism by which LINC01094 regulates CRSwNP progression remains unclear. Our study aimed to investigate the role of LINC01094 in CRSwNP development. METHODS: hNEC were isolated from tissues of controls and CRSwNP patients and stimulated with interleukin (IL)-13. 3-(4, 5-Dimethylthiazolyl2)-2, 5-diphenyltetrazolium bromide (MTT) assay was employed to analyze hNEC viability. Flow cytometry was employed to analyze pyroptosis. Immunofluorescence was employed to analyze Snail nuclear translocation. The interactions between LINC01094, fused in sarcoma (FUS) and high mobility group box-1 (HMGB1) were analyzed by RNA immunoprecipitation (RIP) and RNA pull-down assays. RESULTS: LINC01094 and EMT-related proteins were markedly upregulated in nasal polyp tissues of CRSwNP. LINC01094 knockdown inhibited IL-13-induced hNEC EMT and pyroptosis. LINC01094 promoted HMGB1 expression in CRSwNP by binding with FUS. HMGB1 promoted Snail nuclear import in GSK-B phosphorylation-dependent manner. CONCLUSION: LINC01094 facilitated hNEC EMT and pyroptosis in CRSwNP by activating the HMGB1/GSK-B Snail axis, which suggested that LINC01094 might serve as a biomarker and therapeutic target in CRSwNP.

[Research progress on uranium induced human renal injury and its risk prediction].

Uranium has both radiotoxicity and chemical toxicity. Low enriched uranium is mainly chemically toxic, the kidney is the target organ of uranium chemical toxicity. However, due to the differences among species and the mixed effects of chemical toxicity and radiotoxicity, the dose effect relationship of uranium is not clear, and the current standards in China do not provide chemical toxicity limits for uranium workplaces. This paper reviews the data of acute and chronic human uranium exposure, dose effect relationship and renal injury risk prediction literature at home and abroad, providing reference for the health protection of uranium workers and the establishment of chemical limits in uranium workplaces.

[Analysis of noise reduction measures in a noise workshop handover control room].

Objective: To explore the sound insulation, sound absorption and other noise reduction transformation methods in a noise workshop handover control room. Methods: In December 2021, through the occupational health investigation and on-site testing of the handover control room of a noise workshop, the causes of excessive noise were analyzed, and the transformation design scheme to reduce noise was proposed and the effect was analyzed. Results: Before the transformation, the peak frequency band noise intensity of the noise workshop handover control room was 112.8 dB (A), and the peak frequency was 1000 Hz. After noise reduction, the theoretical calculated control value was 61.0 dB (A), and the measured noise intensity was 59.8 dB (A) . Conclusion: The noise intensity of the handover control room is reduced after noise reduction, which is in line with the contact limit requirements of the control room in GBZ 1-2010 "Hygienic Standards for the Design of Industrial Enterprises", and has reference significance for noise control engineering.

[Design for online monitoring of occupational hazard factors based on internet of things].

At present, there are disadvantages with the detection for occupational hazard factors, such as insufficient monitoring data, poor timeliness, weak representativeness, long detection cycles, and inability to continuously monitor. Taking advantages of internet of things technology, an online monitoring platform for occupational hazard factors has been designed. The platform collects the concentration (intensity) of hazard factors through sensors, transmits the occupational hazards data collected online in realtime. The online monitoring cloud center for occupational hazard factors processes and analyzes online monitoring data in realtime, stores the hazard factors data to form database management, and provides user application services to form an intelligent online monitoring service model for occupational hazard factors. Based on the online monitoring platform of occupational hazard factors, multi-level government health supervision departments and employers can grasp the status of hazard factors in real time, which is conducive to improving the level of occupational hazard supervision.

[Kindlin-2 regulates endometrium development via mTOR and Hippo signaling pathways in mice].

OBJECTIVE: To investigate the effects and mechanisms of Kindlin-2 on uterus development and reproductive capacity in female mice. METHODS: Cdh16-Cre tool mice and Kindlin-2flox/flox mice were used to construct the mouse model of uterus specific knockout of Kindlin-2, and the effects of Kindlin-2 deletion on uterine development and reproduction capacity of female mice were observed. High expression and knockdown of Kindlin-2 in endometrial cancer cell lines HEC-1 and Ish were used to detect the regulation of mammalian target of rapamycin (mTOR) signaling pathway. In addition, uterine proteins of the female mice with specific knockout of Kindlin-2 and female mice in the control group were extracted to detect the protein levels of key molecules of mTOR signaling pathway and Hippo signaling pathway. RESULTS: The mouse model of uterine specific knockout of Kindlin-2 was successfully constructed. The knockout efficiency of Kindlin-2 in mouse uterus was identified and verified by mouse tail polymerase chain reaction (PCR), Western blot protein identification, immunohistochemical staining (IHC) and other methods. Compared with the control group, the female mice with uterus specific deletion of Kindlin-2 lost weight, seriously impaired reproductive ability, and the number of newborn mice decreased, but the proportion of the female mice and male mice in the newborn mice did not change. Hematoxylin eosin staining (HE) experiment showed that the endometrium of Kindlin-2 knockout group was incomplete and the thickness of uterine wall became thinner. In terms of mechanism, the deletion of Kindlin-2 in endo-metrial cancer cell lines HEC-1 and Ish could downregulate the protein levels of mTOR, phosphorylated mTOR, adenosine monophosphate-activated protein kinase (AMPK), phosphorylated AMPK and phosphorylated ribosomal protein S6 (S6), and the mTOR signal pathway was inhibited. It was found that the specific deletion of Kindlin-2 could upregulate the protein levels of Mps one binding 1 (MOB1) and phosphorylated Yes-associated protein (YAP) in the uterus of the female mice, and the Hippo signal pathway was activated. CONCLUSION: Kindlin-2 inhibits the development of uterus by inhibiting mTOR signal pathway and activating Hippo signal pathway, thereby inhibiting the fertility of female mice.

[Study on the diffusion of hydrogen fluoride in an electrolytic fluoride plant based on CFD numerical simulation method].

Objective: Using CFD technology to grasp the distribution and diffusion of hydrogen fluoride in an electrolytic fluorine plant, provide guidance and scientific basis for enterprises to carry out occupational health management in enterprises, install hazardous substance alarm devices, and protect workers' occupational health. Methods: In July 2019, the diffusion law of hydrogen fluoride gas produced in an electrolytic fluorine plant is selected as the research object. Through the establishment of models and grids, the Fluent numerical simulation method is finally used to simulate the diffusion and distribution of hydrogen fluoride gas under ventilation conditions. Results: The results showed that the average concentration of hydrogen fluoride was 0.045 mg/m(3) in the workplace, and the absorbed zone height (1.5 m) was 0.02 mg/m(3) in the inspection channel, which was in accordance with the national standard. However, there is eddy current above the electrolyzer near the inlet, may lead to the accumulation of hydrogen fluoride gas. Conclusion: The research of CFD numerical simulation method on the distribution and diffusion of hydrogen fluoride concentration in electrolytic fluorine plant can be applied to the prevention, control and management of occupational hazards in electrolytic fluorine plant.

[Application status of CFD technology in occupational hazard control].

In the process of occupational hazard management, computational fluid dynamics technology can be used to reflect the distribution pattern of occupational hazards in the production process, so as to quickly and accurately guide the formulation of occupational disease prevention and control programs. This paper summarizes and analyzes the current research results on the prevention and control of occupational hazards in workplaces through computational fluid dynamics technology, and describes the application of these research results in the process of occupational disease prevention and control. On this basis, this paper presents the problems and application limitations of existing research and points out the future key research directions, which are of great reference value for guiding further systematic and in-depth research on simulation, experimentation and management of occupational hazards that can cause occupational diseases.

Machine learning-based radiomic evaluation of treatment response prediction in glioblastoma.

AIM: To investigate machine learning based models combining clinical, radiomic, and molecular information to distinguish between early true progression (tPD) and pseudoprogression (psPD) in patients with glioblastoma. MATERIALS AND METHODS: A retrospective analysis was undertaken of 76 patients (46 tPD, 30 psPD) with early enhancing disease following chemoradiotherapy for glioblastoma. Outcome was determined on follow-up until 6 months post-chemoradiotherapy. Models comprised clinical characteristics, O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation status, and 307 quantitative imaging features extracted from enhancing disease and perilesional oedema masks on early post-chemoradiotherapy contrast-enhanced T1-weighted imaging, T2-weighted imaging (T2WI), and apparent diffusion coefficient (ADC) maps. Feature selection was performed within bootstrapped cross-validated recursive feature elimination with a random forest algorithm. Naive Bayes five-fold cross-validation was used to validate the final model. RESULTS: Top selected features included age, MGMT promoter methylation status, two shape-based features from the enhancing disease mask, three radiomic features from the enhancing disease mask on ADC, and one radiomic feature from the perilesional oedema mask on T2WI. The final model had an area under the receiver operating characteristics curve (AUC) of 0.80, sensitivity 78.2%, specificity 66.7%, and accuracy of 73.7%. CONCLUSION: Incorporating a machine learning-based approach using quantitative radiomic features from standard-of-care magnetic resonance imaging (MRI), in combination with clinical characteristics and MGMT promoter methylation status has a complementary effect and improves model performance for early prediction of glioblastoma treatment response.

Standard diffusion-weighted, intravoxel incoherent motion, and dynamic contrast-enhanced MRI of musculoskeletal tumours: correlations with Ki67 proliferation status.

AIM: To determine whether quantitative parameters derived from conventional diffusion-weighted imaging (DWI), intravoxel incoherent motion (IVIM), and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) correlate with the Ki67 proliferation status in musculoskeletal tumours. MATERIALS AND METHODS: Twenty-eight patients with musculoskeletal tumours diagnosed via surgical specimen histological analysis who underwent standard DWI, IVIM, and DCE were reviewed retrospectively. The mean standard DWI (apparent diffusion coefficient [ADC]), IVIM (pure diffusion coefficient [D], pseudo-diffusion coefficient [D∗] and perfusion fraction [ƒ]), and DCE (volume transfer constant [Ktrans], rate constant [Kep], and extravascular extracellular volume fraction [Ve]) parameters were measured and correlated with the Ki67 index. The Ki67 value was categorised as high (>20%) or low (≤20%). RESULTS: The ADC and D values correlated negatively with the Ki67 index (r=-0.711∼-0.699, p<0.001), whereas the Ktrans and Kep values correlated positively with the Ki67 index (r=0.389-0.434, p=0.021, 0.041). The ADC and D values were lower (p<0.001), whereas the Ktrans and Kep values were higher (p=0.011, 0.005) in musculoskeletal tumours with a high Ki67 status than in those in a low status. The ADC and D demonstrated the largest area under the receiver-operating characteristic curve (AUC = 0.953), which is statistically bigger than the AUC of Ktrans and Kep (0.784 and 0.802, respectively). CONCLUSION: ADC, D, Ktrans, and Kep correlate with the Ki67 index. ADC and D are the strongest quantitative parameters for predicting Ki67 status.

Inhibition of carbonic anhydrase II by sulfonamide derivatives.

A series of sulfonamide derivatives were synthesized, and the enzyme inhibitory activity of the synthesized compounds on carbonic anhydrase II was evaluated. Through molecular docking studies, it was found that compounds 1b, 1e, 2a, 2b, 3a have a strong binding affinity to carbonic anhydrase II. The IC50 values of the four compounds 1e, 2b, 3a, and 3b were lower than that of the positive control drug acetazolamide. What's more, the compounds had a high inhibitory activity for A549 lung cancer cell growth, among them, 1e and 3a could inhibit both carbonic anhydrase II and lung cancer cell proliferation.

Operating behavior of micro-LEDs on a GaN substrate at ultrahigh injection current densities.

Near-ultraviolet micro-LEDs with different diameters were fabricated on GaN substrates. The electroluminescence and the light output power-current density and current density-voltage relationships were measured. A saturated current density of 358 kA/cm2 was achieved with a 20 µm LED. The ideality factor curves showed steps and peaks when the injection current density was increased from 20 to 150 kA/cm2 and an abnormal efficiency increase. The transport and recombination processes of micro-LEDs at high injection current densities were simulated, and the many-body effect and phase space filling in the integrated quantum drift-diffusion model were considered. Serious current crowding was observed above 100 kA/cm2, even for the 20 µm LED.

Multi-modular engineering for renewable production of isoprene via mevalonate pathway in Escherichia coli.

AIMS: To establish the biotechnology platforms for production of bio-based chemicals in various micro-organisms is considered as a promising target to improve renewable production of isoprene. METHODS AND RESULTS: In this study, we heterologously expressed the mevalonate (MVA) isoprene biosynthesis pathway, and explored three strategies of increasing isoprene production in Escherichia coli. We first manipulated the expression levels of the MVA pathway genes through changing the gene cassettes and promoters. To introduce cofactor engineering, we then overexpressed NADP-dependent glyceraldehyde-3-phosphate dehydrogenase gene from Clostridium acetobutylicum to supply available NADPH. To reduce the inhibitory by-product accumulation, we finally knocked out acetate-producing genes, phosphate acetyl transferase and pyruvate oxidase B in E. coliJM109 (DE3), decreasing acetate accumulation 89% and increasing isoprene production 39%. The strategies described here finally increased the isoprene titre to 92 mg l-1 in two-gene deletion strain JMAB-4T7P1Trc, increasing 2·6-fold comparing to strain JM7T7. CONCLUSION: The multimodularly engineering approaches including promoter engineering, cofactor engineering and by-product reducing could be used to improve isoprene production in E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: The metabolic strategies in this study show us directions for further studies to promote transformation of renewable sources to isoprene.

[Analysis of infection status and recombination types of norovirus in patients with acute gastroenteritis in the Ningxia Hui Autonomous Region from 2016 to 2017].

Objective: To analyze the infection status and recombination of Norovirus in patients with acute gastroenteritis in Ningxia. Methods: The specimens of 10 sentinel hospitals in Ningxia were collected from 2016 to 2017. Real-time quantitative PCR was used for nucleic acid detection. GⅡ-positive samples were amplified by RT-PCR for the RdRp and Capsid regions, then sequenced and genotyped. Evolution analysis was performed using software such as MEGA-X, and recombination analysis was performed using Simplot 3.5.1 and RDP4. Results: The age of the 2 334 cases was 1.42 (0.68, 7.69) years old, 1 133 cases in 2016 and 1 201 cases in 2017, 1 343 and 991 cases for males and females respectively. The positive rate of Norovirus GⅠ genogroup was 0.86% (20/2 334), and GⅡ genogroup was 14.82% (346/2 334). A total of 78 recombinant strains were sequenced and 12 recombinant types were found. GⅡ.Pe/GⅡ.4Sydney_2012 and GⅡ.P12/GⅡ.3 were the main epidemic strains, accounting for 35.90% (28 strains) and 32.05% (25 strain) respectively, followed by GⅡ.P16/GⅡ.2 accounting for 12.82% (10 strains). Among them,GⅡ.P7/GⅡ.6 (2 strains), GⅡ.P12/GⅡ.3 (6 strains), GⅡ.P16/GⅡ.1 (2 strains), GⅡ.P16/GⅡ.2 (5 strains), GⅡ.Pe/GⅡ.4 (7 strains) were detected for the first time in Ningxia. Recombinant strains were all intergenotype recombination, and the recombination breakpionts were all located within ORF1. Conclusion: Norovirus infection in Ningxia area was mainly in GⅡ genogroup from 2016 to 2017, and most of them were recombinant strains. GⅡ.Pe/GⅡ.4Sydney_2012 and GⅡ.P12/GⅡ.3 were the main epidemic strains, followed by GⅡ.P16/GⅡ. 2.

[Pre-descement membrane endothelial keratoplasty for treatment of patients with corneal endothelial decompensation].

Objective: To investigate the clinical outcomes of Pre-descement membrane endothelial keratoplasty (PDEK) for treatment of patients with corneal endothelial decompensation. Methods: Retrospective study of case series. The cases of 20 patients (20 eyes) who were diagnosed with corneal endothelial decompensation induced by various original diseases and received PDEK during July 2016 and December 2016 at Zhongshan Ophthalmic Center have been analyzed. The participants included 8 males and 12 females with an average age of (59.3±11.8) years. All 20 patients received PDEK operation, the Dua's layer, descement membrane and endothelium of donor tissue were separated from the underlying stroma with the application of big-bubble technique, and donor discs with diameters of 7.75 or 8.00 mm were harvested with scissors. The donor discs were stained with 0.06% trypan blue and loaded on inserters which were then gently inserted into the anterior chamber of the recipient. When the rolled inserted graft was unfolded, air was carefully injected into the anterior chamber underneath the graft so that the graft can attach to the stroma of the recipient. Post-operation follow-ups of over 6 months have been conducted for all patients, the best spectacle corrected visual acuity (BSCVA), the position of donor disc, corneal thickness and corneal endothelial cell density (ECD) were documented. The pre-operation and post-operation (6 months postoperatively) corneal thickness data of the patients were analyzed with paired sample t test. Results: The success rate of preparing PDMEK donor disc with big-bubble technique is 90% (18/20). Eighteen patients (90%) received PDMEK surgery successfully. Anterior segment optical coherence tomography (AS-OCT) results indicated that sixteen donor discs (16/18) were well attached to the back surface of the recipient stroma, and that two discs (2/18) dislocated at 6 days after surgery. At 6 months post-operatively, the corneas of 14 patients (14/18) turned clear with their BSCVA ranging 0.4 to 1.0, and the ECD was (1 389.3±373.2) cells/mm(2) for the patients with clear corneas. At 6 months post-operatively, the average corneal thickness of the patients reduced to (605±45) µm from the preoperative level of (655±56) µm, and the differences are of statistical significance (t=2.137, P=0.032). Conclusions: Application of big-bubble technique could effectively secure the success rate of PDEK disc preparation and control the loss of donated corneas. PDEK disc can be easily handled and unrolled in the anterior chamber, which could improve the postoperative clinical outcomes. (Chin J Ophthalmol, 2018, 54: 105-110).

[Descemet's membrane endothelial keratoplasty for treatment of patients with corneal endothelial decompensation].

Objective: To evaluate the clinical outcomes of Descemet's membrane endothelial keratoplasty (DMEK) for treatment of patients with corneal endothelial decompensation in China. Methods: Interventional case series. Ten patients (10 eyes) with the diagnosis of corneal endothelial decompensation received DMEK during July 2016 and September 2016 in Zhongshan Ophthalmic Center. Descemet's membrane of the donor tissue was carefully stripped from the underlying stroma manually and cut with a 7.75- or 8.0-mm-diameter donor punch. After stained with 0.06% typan blue, the donor disc loaded on an inserter was gently inserted into the anterior chamber of the recipient. When the rolled graft was unfolded, air was carefully injected into the anterior chamber underneath the graft, so that the graft attached to the stroma of the recipient. All patients were followed up for 3-6 months. The best spectacle corrected visual acuity, the status of donor disc, corneal thickness and corneal endothelial cell density were documented and analyzed. Results: At 1 day after surgery, 1 donor disc presented a rolled edge, and repeated DMEK surgery was performed for this patient. The remaining 9 donor discs presented in place well. At 1 month after surgery, 7 donor discs were found to well attach to the back surface of the recipient stroma with normal corneal posterior surface using anterior segment optical coherence tomography. The average corneal thickness reduced to (620±67) µm from preoperative (680±78.3) µm (t=2.677, P=0.025). At 3 months after surgery, 6 patients achieved clear corneas with best spectacle corrected visual acuity ranging from 0.4 to 0.8. The endothelial cell density was 1 342.3±436.5 (872 to 1 933) cells/mm(2) in the patients with clear corneas. The average corneal thickness reduced to (612±70) µm (t=2.971, P=0.016). Conclusions: DMEK could improve visual acuity for corneal endothelial decompensation, but there were more technical difficulties in Chinese eyes. (Chin J Ophthalmol, 2017, 53: 534-539).

Adaptive behaviour of Chinese boys with fragile X syndrome.

BACKGROUND: Adaptive behaviour is closely related to quality of life in children with intellectual disability (ID), but little is known about the adaptive behaviour of children with fragile X syndrome (FXS) in China. METHOD: In boys with FXS, the adaptive behaviours in six domains, including self-dependence, locomotion, work skills, communication, socialisation and self-management, were assessed by the Infants-Junior Middle School Students Social-life Abilities Scale. In addition, we compared the adaptive skills of boys with FXS to those of three control groups of boys, including boys with Down syndrome (DS) and typically developing (TD) boys matched by chronological age (CA) or mental age (MA). The profile of the adaptive behaviour of boys with FXS is discussed in detail. RESULTS: Compared to boys with DS, boys with FXS obtained lower scores in three domains in adaptive behaviour, including work skills, socialisation and self-management skills; boys with FXS had better scores in self-dependence and locomotion skills than boys matched for MA; as expected, boys with FXS had significantly poorer adaptive skills in all six domains assessed compared to CA boys. CONCLUSION: The development of adaptive skills in boys with FXS was worse than that of boys with DS. The profile of the adaptive behaviour of boys with FXS establishes a basis for the development of targeted interventions to promote social development in this population.

Elevated HOXB9 expression promotes differentiation and predicts a favourable outcome in colon adenocarcinoma patients.

BACKGROUND: Little is known about the tumour suppressive proteins and the underlying mechanisms that suppress colon cancer progression. Homeodomain-containing transcription factor HOXB9 plays an important role in embryogenesis and cancer development. We here aim to uncover the potential role of HOXB9 in the regulation of colon adenocarcinoma progression including epithelial-to-mesenchymal transition. METHODS: HOXB9 expression in colon adenocarcinoma cells and patients was analysed by western blot and immunohistochemistry separately. Correlation between HOXB9 expressions with patients' survival was assessed by Kaplan-Meier analysis. HOXB9-regulated target gene expression was determined by RNA sequencing in HOXB9-overexpressing colon adenocarcinoma cells. RESULTS: Elevated HOXB9 expression was identified in well-differentiated colon adenocarcinoma patients and was associated with a better overall patients' survival. Overexpression of HOXB9 inhibited colon adenocarcinoma cell growth, migration, invasion in vitro and tumour growth, liver as well as lung metastases in nude mice; whereas silencing HOXB9 promoted these functions. HOXB9 promoted colon adenocarcinoma differentiation via a mechanism that stimulates mesenchymal-to-epithelial transition, involving downregulation of EMT-promoting transcriptional factors including Snail, Twist, FOXC2 and ZEB1 and upregulation of epithelial proteins including E-cadherin, claudins-1, -4, -7, occludin and ZO-1. CONCLUSIONS: HOXB9 is a novel tumour suppressor that inhibits colon adenocarcinoma progression by inducing differentiation. Elevated expression of HOXB9 predicts a longer survival in colon adenocarcinoma patients.

Multiple-trait genome-wide association study based on principal component analysis for residual covariance matrix.

Given the drawbacks of implementing multivariate analysis for mapping multiple traits in genome-wide association study (GWAS), principal component analysis (PCA) has been widely used to generate independent 'super traits' from the original multivariate phenotypic traits for the univariate analysis. However, parameter estimates in this framework may not be the same as those from the joint analysis of all traits, leading to spurious linkage results. In this paper, we propose to perform the PCA for residual covariance matrix instead of the phenotypical covariance matrix, based on which multiple traits are transformed to a group of pseudo principal components. The PCA for residual covariance matrix allows analyzing each pseudo principal component separately. In addition, all parameter estimates are equivalent to those obtained from the joint multivariate analysis under a linear transformation. However, a fast least absolute shrinkage and selection operator (LASSO) for estimating the sparse oversaturated genetic model greatly reduces the computational costs of this procedure. Extensive simulations show statistical and computational efficiencies of the proposed method. We illustrate this method in a GWAS for 20 slaughtering traits and meat quality traits in beef cattle.

Gene cloning of porcine adiponectin gene from adipose tissue and construction of its eukaryotic expression vector.

To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18-T vector to construct recombinant clonal vector pMD-ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA-ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA-ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 µg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA-ADPN constructed in this study effectively.

First Report of Pineapple Heart Rot Caused by Phytophthora nicotianae in Hainan Province, China.

Pineapple (Ananas comosus) is an economically important tropical fruit in Hainan Province, China. During September to November 2011, heart rot disese of pineapple was found in Ledong and Wangning of Hainan Province. A survey of 150 ha producing areas of pineapple revealed that the fields were affected at an incidence ranging from 25% to 30%. Infected plants showed water-soaked lesions and soft rot on the base of heart leaves near the soil surface. Heart leaves of infected plants were easily pulled out. As the disease progressed, plants collapsed and died. Diseased tissue fragments (2 × 2 mm) were surface-disinfected for 10 min with 0.3% NaClO, then rinsed three times in sterile water, and plated to 10% V8 juice agar (4). Inoculated dishes were incubated at 26°C in the dark. After 5 days, Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were isolated from the tissue cultures, which has aseptate hyphae. Sporangia were papillate, noncaducous, oval or spherical, and 34.5 to 58.2 µm. Clamydospores, both terminal and intercalary, were also spherical, and were 23.4 to 34.0 µm (2). The ITS region of rDNA was amplified using primers ITS4/ITS5, and the 927-bp product of the ITS showed 99% sequence identity to Phytophthora nicotianae (GenBank Accession No. JF792540), and the sequence was accessed to NCBI (JX978446). Pathogenicity tests were confirmed by irrigating the wounded stem bases of 10 2-month-old pineapple plants with 50 ml of P. nicotionae zoospore solution (15,000 zoospores/ml), and another 10 plants of the same cultivar inoculated with sterile water served as controls. Plants were placed in pots in a greenhouse at 28°C and 90% relative humidity. After 9 days, soft rot was observed clearly on the base of heart leaves of all 10 inoculated plants, while the control plants appeared normal. P. nicotianae was reisolated from the infected pineapple plants, and confirmed to be the same as the inoculated pathogen by conducting a ITS rDNA sequence comparison and morphological characteristics. P. nicotianae was previously reported as the causal agent of heart rot of pineapple in Hawaii, U.S.A. (3) and Guangdong Province of China (1). To our knowledge, this is the first report of P. nicotianae on pineapple in Hainan Province, China. References: (1) J. Z. Chen et al. J. Yunnan Agric. Univ. 8:134, 2003. (2) H. H. Ho. Mycologia 73:705, 1981.(3) K. W. Howard et al. Plant Dis. Rep. 48:848, 1964. (4) X. B. Zheng. Page 81 in: Phytophthora and its Research Technology. China Agricultural Press, Beijing, 1997.